ObjectiveThe U6 promoter is an essential element for driving sgRNA expression in the clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein 9（CRISPR/Cas9）gene editing system in dicotyledonous plants. Endogenous U6 promoters typically exhibit higher transcriptional activity， which can significantly improve gene editing efficiency. This study aims to identify endogenous U6 promoters in Artemisia annua to optimize its CRISPR/Cas9 gene editing system， which holds significant importance for its molecular breeding. MethodsOn the basis of the highly conserved U6 snRNA sequences in Arabidopsis thaliana， endogenous U6 promoters were screened in the A. annua genome. Expression vectors were constructed with candidate AaU6 promoter driving the firefly luciferase （LUC） reporter gene， and then transiently transformed into Nicotiana benthamiana. Transcriptional activities of the promoters were measured and compared by in vivo imaging and the Dual Luciferase Reporter assay. ResultsEight endogenous U6 promoters were successfully cloned from A. annua. Sequences alignment revealed that all these promoters contained the two conserved cis-acting elements， upstream sequence element （USE） and TATA-box， which affected their transcriptional activity. Dual-luciferase activity assays indicated that the transcriptional activities of AaU6-3， AaU6-1， and AaU6-5 were significantly higher than that of the Arabidopsis AtU6-26 promoter， with AaU6-3 exhibiting the highest activity. ConclusionThis study identified three endogenous AaU6 promoters with high transcriptional activity in A. annua， providing key functional elements for establishing an efficient gene editing system in A. annua. These findings will contribute to advancing precision molecular breeding and high-quality germplasm innovation in A. annua.

Glioma represents the most prevalent malignant tumor of the central nervous system, with chemotherapy serving as an essential adjunctive treatment. However, most chemotherapeutic agents exhibit limited ability to penetrate the blood-brain barrier (BBB). This study introduced a novel dual-targeting strategy for glioma therapy by modulating the formation of nanobody-driven protein coronas to enhance the brain and tumor-targeting efficiency of hydrophobic cisplatin prodrug-loaded lipid nanoparticles (C8Pt-Ls). Specifically, nanobodies (Nbs) with fibrinogen-binding capabilities were conjugated to the surface of C8Pt-Ls, resulting in the generation of Nb-C8Pt-Ls. Within the bloodstream, Nb-C8Pt-Ls could bound more fibrinogen, forming the protein corona that specifically interacted with LRP-1, a receptor highly expressed on the BBB. This interaction enabled a "Hitchhiking Effect" mechanism, facilitating efficient trans-BBB transport and promoting effective brain targeting. Additionally, the protein corona interacted with LRP-1, which is also overexpressed in glioma cells, achieving precise tumor targeting. Computational simulations and SPR detection clarified the molecular interaction mechanism of the Nb-fibrinogen-(LRP-1) complex, confirming its binding specificity and stability. Our results demonstrated that this strategy significantly enhanced C8Pt accumulation in brain tissues and tumors, induced apoptosis in glioma cells, and improved therapeutic efficacy. This study provides a novel framework for glioma therapy and underscores the potential of protein corona modulation-based dual-targeting strategies in advancing treatments for brain tumors.

Objectives:To evaluate the cost-effectiveness of typical pharmaceutical smoking cessation intervention strategies in China in the context of primary cancer prevention.Methods:Markov cohort simulation models were established to simulate the burden of 12 smoking caused cancer, including lung cancer, oral cancer, nasopharyngeal cancer, laryngeal cancer, esophageal cancer, gastric cancer, pancreatic cancer, liver cancer, kidney cancer, bladder cancer, cervical cancer, and acute myeloid leukemia. Taking incremental cost effectiveness ratio (ICER) as the main indicator, the model sets one year as the cycling period for 50 periods and simulates the cohort of 10 000 thirty-five-year-old current smokers with various smoking cessation strategies. To ensure the robustness of conclusion, univariate sensitivity analysis, probability sensitivity analysis, and age-group sensitivity analysis were conducted.Results:The results showed that varenicline intervention was the most cost-effective intervention. Compared to the next most effective option, incremental cost of each additional quality-adjusted life year is 11 140.28 yuan, which is below the threshold of willingness to pay (1 year GDP per capita). The value of ICER increased as the increasing age group of adopting intervention, but neither exceeded the threshold of willingness to pay. One-way sensitivity analysis showed that the value of discount rate, the hazard ratio and cost of intervention strategy had a greater impact on the result of ICER.Conclusion:In China, the use of varenicline to quit smoking is highly cost effective in the context of cancer primary prevention, especially for younger smokers.

Alzheimer's disease(AD)is the most common form of senile dementia,and its pathogenesis is still un-clear.While β-amyloid(Aβ)is considered an important cause of AD,the pathological mechanism of Aβ inducing AD is subject to various controversies.Recent studies have shown that the myeloid cell trigger receptor(TREM2)plays an impor-tant role in the pathological process of AD,and it can not only serve as an important receptor for the internalization of Aβ but also become a biological diagnostic biomarker and therapeutic target.Hence,elucidating the structure and function of TREM2 will provide important ideas for the prevention and treatment of AD.This article will provide a systematic review of the structure of TREM2,its impact on microglial cell function,its pathological role in AD,and the current status of target-ed TREM2 therapy for AD.These summaries will provide valuable references for basic research on AD.

Objective To observe the role of dried tangerine peel in Yigong Powder improves iron metabolism and promotes red blood cell generation in anemia of chronic disease (ACD).Methods With a two-by-two factorial design,the Yigong Powder was divided into dried tangerine peel and Chenpi absent Decoction. According to the random number table method,32 zymosan-induced generalized inflammation (ZIGI) mice were randomly divided into the model group,the dried tangerine peel group,the Chenpi absent Decoction group,and the Yigong Powder group. The dried tangerine peel group,Chenpi absent Decoction group and the Yigong Powder group were given dried tangerine peel(3.083 g/kg),Chenpi absent Decoction(12.33g/kg),and Yigong Powder(15.413g/kg)by gavage to the corresponding group of mice. The model group was given an equal amount of physiological saline by gavage,and treated continuously for 7 days. After the completion of administration,the body weight of each group of mice was recorded. The hemoglobin content of each group of mice was detected using a fully automatic cell counter,the serum iron content was detected using colorimetry,the serum ferritin content was detected using enzyme-linked immunosorbent assay (ELISA),and the spleen index was calculated. The liver tissue inflammatory factors interleukin-1β (IL-1β),interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),interferon-γ (IFN-γ),interleukin-4 (IL-4),and interleukin-10 (IL-10) levels were detected using Luminex method. The mRNA expressions of liver tissue hepcidin gene (HAMP) and membrane iron transporter ( Fpn) were detected using real-time fluorescence PCR method. Results Dried tangerine peel and Chenpi absent Decoction both showed interactive effects in regulating hemoglobin,serum iron,serum ferritin content,improving spleen index,and regulating the mRNA expressions of HAMP,Fpn,as well as IL-1β and IFN-γ (P<0.05). Compared with the model group,dried tangerine peel significantly increased hemoglobin,serum iron content,and Fpn mRNA expression in ZIGI model mice,while decreasing ferritin content,spleen index,HAMP mRNA expression,and the levels of IL-1β,IL-6,TNF-α,and IFN-γ (P<0.05). Chenpi absent Decoction significantly increased serum iron content and Fpn mRNA expression in ZIGI model mice,while reducing spleen index,ferritin content,HAMP mRNA expression,and the levels of IL-1β and IFN-γ、IL-4 (P<0.05). Conclusion The effects of dried tangerine peel on inflammatory factors (IL-6 and TNF-α) and Fpn may play a key role in the improvement effects of Yigong Powder on ACD and iron metabolism.

Objective:To examine the distribution features of angiotensin converting enzyme ( ACE) gene I/D polymorphisms and their correlation with left ventricular hypertrophy (LVH) in children and adolescents with essential hypertension. Methods:This was a case-control study.One hundred and forty-four children and adolescents who were diagnosed with essential hypertension but received no treatment at the Department of Cardiology, Children′s Hospital, Capital Institute of Pediatrics from January 2022 to April 2023 were recruited. ACE gene polymorphisms were detected using the polymerase chain reaction-restriction fragment length polymorphism method.Patients were divided into the DD+ DI genotype group and the II genotype group according to the dominant model, and the differences in gene frequencies, genotype frequencies, clinical data, and plasma renin-angiotensin-aldosterone system (RAAS) levels were compared between the two groups.In addition, according to the echocardiographic findings, patients were divided into the LVH and non-LVH groups.The risk factors of LVH were analyzed using the multivariate Logistic regression model.The correlation of ACE gene I/D polymorphisms with LVH in children and adolescents with essential hypertension was evaluated using the Spearman analysis.The correlation between plasma RAAS levels and LVH was assessed using the partial correlation analysis. Results:The frequencies of DD, DI and II genotypes in 144 patients were 11.1%, 50.7% and 38.2%, respectively.The frequency of the D and I allele was 36.5%(105/288) and 63.5%(183/288) respectively.Patients in the DD+ DI genotype group (89 cases) were older than those in the II genotype group (55 cases) ( Z=-2.308, P=0.021), and there was no statistically significant difference between the two groups in gender and height (all P>0.05).The incidence of LVH in the DD+ DI genotype group was higher than that in the II genotype group, with a statistically significant difference ( χ2=5.230, P=0.022).Renin activity in RAAS levels was positively correlated with left ventricular mass index and relative wall thickness ( r=0.276, 0.247; P=0.002, 0.006).Multivariate Logistic regression analysis showed that both DD+ DI genotype ( OR=5.678, 95% CI: 1.623-19.872) and body mass index (BMI) ( OR=1.124, 95% CI: 1.024-1.233) were risk factors for the development of LVH in children and adolescents with essential hypertension (all P<0.05). Conclusions:ACE gene I/D polymorphisms are strongly associated with LVH.DD+ DI genotype and BMI are independent risk factors for the development of LVH in children and adolescents with essential hypertension.

Objective:To summarize the content and effects of exercise interventions for stroke survivors, providing references for medical and nursing personnel in formulating exercise intervention plans for these patients.Methods:Literature on exercise interventions for stroke survivors was searched in databases such as China National Knowledge Infrastructure, Wanfang, VIP, China Biology Medicine disc, Cochrane Library, PubMed, Embase, and Web of Science up to May 1, 2023. Data extraction and summarization were conducted on the included literature.Results:Twenty-two articles were included in the review. The exercise types for stroke survivors primarily involved aerobic and resistance training; the intensity was mainly moderate to high; the frequency was mostly 3-5 times per week, with each session lasting 30-60 minutes; and the duration of interventions ranged from as short as two weeks to as long as one year. Exercise interventions improved stroke survivors' functional mobility, quality of life, cardiopulmonary function, mental state, cognitive function, fatigue, sleep, and compliance to varying degrees.Conclusions:Exercise interventions have achieved positive outcomes in stroke survivors. In the future, medical and nursing personnel can incorporate patient preferences into exercise intervention plans while ensuring patient safety to promote rehabilitation.

Objective:To conduct an overview of systematic reviews on the safety and effectiveness of early mobilization in stroke patients.Methods:Systematic reviews of early mobilization in stroke were electronically searched on China National Knowledge Infrastructure, WanFang Data, VIP, Chinese Biology Medicine, Cochrane Library, PubMed, Web of Science, CINAHL, Embase, and the Joanna Briggs Institute Evidence-Based Health Care Center Database. The search period was from database establishment to May 2023. Two researchers independently conducted literature screening, data extraction, and methodological quality evaluation of the included literature, as well as evidence summary and grading.Results:A total of 10 systematic reviews were included. The methodological quality evaluation showed that one article was of medium quality, two articles were of low quality, and seven articles were of extremely low quality. The evidence quality evaluation indicated that there were three pieces of high-quality evidence, 17 pieces of medium-quality evidence, 21 pieces of low-quality evidence, and 17 pieces of extremely low-quality evidence.Conclusions:Early mobilization is beneficial for improving the daily living ability and limb motor function of stroke patients, reducing the incidence of complications, shortening hospital stay, and enhancing their walking ability in the short term. However, the quality of evidence in the systematic review is relatively low, and further improvement is needed in the research design of early mobilization intervention measures to provide strong support for clinical practice.

Objective:To investigate the correlation between the level of treponema pallidum specific antibodies(TPAb)and immunoglobulin A/E/M/G(IgA/E/M/G)and IgG subtypes in active syphilis patients,so as to provide a basis for the comprehensive di-agnosis and treatment and prognosis evaluation of clinical syphilis.Methods:Active syphilis patients admitted to the First Affiliated Hospital of Soochow University in 2023 were divided into 1∶4+and 1∶16+groups according to the syphilis toluidine red unheated se-rum test(TRUST),and healthy control group were selected for the detection of TPAb by chemiluminescence microparticle immunoas-say(CMIA).The contents of IgA/E/M/G and IgG1/2/3/4 were determined by immunoturbidimetry.Results:A total of 206 patients with active syphilis(125 males and 81 females)aged(46.05±19.58)years were included in this study.Compared with the healthy control group,IgG2 was decreased and TPAb,IgA/E/M/G and IgG1/3/4 levels were increased in the active syphilis group,with statistically significant differences in TPAb,IgE and IgG1/2/3 levels(P<0.05).Compared with the healthy control group,TRUST 1∶4+group had statistical differences in TPAb,IgE and IgG2,and TPAb was significantly correlated with IgE and IgG2/4.The levels of TPAb,IgA/E/M/G and IgG1/2/3 in TRUST 1∶16+group were statistically significant,and TPAb was significantly correlated with IgE and IgG4(P<0.05).In addition,there were significant differences in TPAb,IgG and IgG1/3 between the two positive groups(P<0.05).Conclu-sion:The levels of TPAb and Ig in active syphilis patients with different TRUST titers are significantly different.TPAb combined with Ig can be used to monitor the strength of syphilis activity,IgA/M/G and IgG1/3 may be candidate biomarkers.

Objective To systematically evaluate the qualitative research on the experience and feeling of elderly patients with hip fracture during rehabilitation.Methods We searched databases including the PubMed,Embase,Web of Science,Cochrane Library,CNAIHL,CNKI,Wanfang,VIP,SinoMed.The literature of qualitative research on the exercise rehabilitation experience of stroke patients from the establishment of the database to August 17,2023 was retrieved.Results A total of 59 clear research topics were extracted,and 14 new categories were formed after induction and integration,which were finally merged into 5 integrated results.Integration results 1:Perceived benefits of exercise rehabilitation.Integration results 2:Multiple driving forces of exercise rehabilitation.Integration results 3:Gradual loss of patience in long-term rehabilitation.Integrated results 4:Generating new rehabilitation needs and actively coping with them.Integrated results 5:There are many difficulties in carrying out self-rehabilitation.Conclusion During the process of exercise rehabilitation for stroke patients,medical personnel should pay attention to the patients'actual feelings,inner needs,and the practical challenges they face in participating in exercise rehabilitation.Personalized exercise rehabilitation facilitation strategies should be developed for stroke patients,aiming for the scientific and precise advancement of exercise rehabilitation,and ultimately improving long-term rehabilitation outcomes for patients.